Membrane‐Surface Anchoring of Charged Diacylglycerol‐Lactones Correlates with Biological Activities

Synthetic diacylglycerol‐lactones (DAG‐lactones) are effective modulators of critical cellular signaling pathways, downstream of the lipophilic second messenger diacylglycerol, that activate a host of protein kinase C (PKC) isozymes and other nonkinase proteins that share similar C1 membrane‐targeting domains with PKC. A fundamental determinant of the biological activity of these amphiphilic molecules is the nature of their interactions with cellular membranes. This study examines the biological properties of charged DAG‐lactones exhibiting different alkyl groups attached to the heterocyclic nitrogen of an α‐pyridylalkylidene chain, and particularly the relationship between membrane interactions of the substituted DAG‐lactones and their respective biological activities. Our results suggest that bilayer interface localization of the N‐alkyl chain in the R² position of the DAG‐lactones inhibits translocation of PKC isoenzymes onto the cellular membrane. However, the orientation of a branched alkyl chain at the bilayer surface facilitates PKC binding and translocation. This investigation emphasizes that bilayer localization of the aromatic side residues of positively charged DAG‐lactone derivatives play a central role in determining biological activity, and that this factor contributes to the diversity of biological actions of these synthetic biomimetic ligands.     

A selective assay to detect chitin and biologically active nano-machineries for chitin-biosynthesis with their intrinsic chitin-synthase molecules

A new assay system for chitin has been developed. It comprises the chitin-binding protein ChbB in fusion with a His-tag as well as with a Strep-tag, the latter of which was chemically coupled to horseradish peroxidase. With the resulting complex, minimal quantities of chitin are photometrically detectable. In addition, the assay allows rapid scoring of the activity of chitin-synthases. As a result, a refined procedure for the rapid purification of yeast chitosomes (nano-machineries for chitin biosynthesis) has been established. Immuno-electronmicroscopical studies of purified chitosomes, gained from a yeast strain carrying a chitin-synthase gene fused to that for GFP (green-fluorescence protein), has led to the in situ localization of chitin-synthase-GFP molecules within chitosomes.     

Crystallisation of Wild-Type and Variant Forms of a Recombinant Plant Enzyme ��-d-Glucan Glucohydrolase from Barley (Hordeum vulgare L.) and Preliminary X-ray Analysis

Wild-type and variant crystals of a recombinant enzyme β-d-glucan glucohydrolase from barley (Hordeum vulgare L.) were obtained by macroseeding and cross-seeding with microcrystals obtained from native plant protein. Crystals grew to dimensions of up to 500 × 250 × 375 μm at 277 K in the hanging-drops by vapour-diffusion. Further, the conditions are described that yielded the wild-type crystals with dimensions of 80 × 40 × 60 μm by self-nucleation vapour-diffusion in sitting-drops at 281 K. The wild-type and recombinant crystals prepared by seeding techniques achived full size within 5–14 days, while the wild-type crystals grown by self-nucleation appeared after 30 days and reached their maximum size after another two months. Both the wild-type and recombinant variant crystals, the latter altered in the key catalytic and substrate-binding residues Glu220, Trp434 and Arg158/Glu161 belonged to the P4₃2₁2 tetragonal space group, i.e., the space group of the native microcrystals was retained in the newly grown recombinant crystals. The crystals diffracted beyond 1.57–1.95 Å and the cell dimensions were between a = b = 99.2–100.8 Å and c = 183.2–183.6 Å. With one molecule in the asymmetric unit, the calculated Matthews coefficients were between 3.4–3.5 ų·Da⁻¹ and the solvent contents varied between 63.4% and 64.5%. The macroseeding and cross-seeding techniques are advantageous, where a limited amount of variant proteins precludes screening of crystallisation conditions, or where variant proteins could not be crystallized.     

SwarmDock and the Use of Normal Modes in Protein-Protein Docking

Here is presented an investigation of the use of normal modes in protein-protein docking, both in theory and in practice. Upper limits of the ability of normal modes to capture the unbound to bound conformational change are calculated on a large test set, with particular focus on the binding interface, the subset of residues from which the binding energy is calculated. Further, the SwarmDock algorithm is presented, to demonstrate that the modelling of conformational change as a linear combination of normal modes is an effective method of modelling flexibility in protein-protein docking.     

Different Effects of Water and Glycerol on Morphology and Properties of Poly(lactic acid)/Soy Protein Concentrate Blends

When containing extra water, soy protein concentrate (SPC) can behave like a plastic melt rather than particulate filler in blending process. In this study, blends of formulated SPC and poly(lactic acid) (PLA) were prepared. The effects of glycerol and water added to SPC on phase morphology, rheological, thermal, thermal‐dynamic, and mechanical properties of the blends were investigated. The results demonstrated that water was more effective in gelating SPC and transforming it into a plastic than glycerol. With extra water added, SPC in the resulting blend existed as stretched threads with certain degree of interconnectivity between the threads. In contrast, with only glycerol added, SPC existed as large SPC agglomerates. Consequently, the blends made from extra water‐containing SPC exhibited higher mechanical, thermal‐dynamic, and rheological properties.

     

Advances and challenges in protein-ligand docking

Molecular docking is a widely-used computational tool for the study of molecular recognition, which aims to predict the binding mode and binding affinity of a complex formed by two or more constituent molecules with known structures. An important type of molecular docking is protein-ligand docking because of its therapeutic applications in modern structure-based drug design. Here, we review the recent advances of protein flexibility, ligand sampling, and scoring functions-the three important aspects in protein-ligand docking. Challenges and possible future directions are discussed in the Conclusion.     

Effect of oxidative damage due to excessive protein ingestion on pancreas function in mice

The present study was undertaken to evaluate the effect of oxidative damage due to excessive protein diet on pancreas function in mice. For this purpose, thirty male (C57BL/6J) mice were randomly divided into three groups and fed on different diets as follows: group 1 was fed on a normal diet, group 2 was fed on an excessive protein diet and group 3 was fed on an excessive protein diet supplemented with 0.06 g/kg cysteamine. Each group was fed for 2 weeks, and then pancreas samples were collected to examine oxidative and antioxidant parameters and pancreas function. The results showed that ingestion of an excessive protein diet markedly increased contents of malondialdehyde (MDA) and decreased T-AOC and activities of antioxidants SOD and GSH-Px, compared with a normal diet (P < 0.05). Pancreas weight and concentration of protein, DNA and RNA were significantly higher (P < 0.05), digestive enzyme activities were significantly lower and levels of somatostatin and insulin were higher in mice fed with an excessive protein diet than those fed with a normal protein diet. In the group fed with excessive protein diet supplemented with cysteamine, oxidative stress was mitigated and pancreas function was improved. These data demonstrate that excessive protein ingestion could increase oxidative damage of free radicals on pancreas function through destroying the balance of oxidants and antioxidants.     

戊二醛接枝共聚改性骨胶粘合剂的研究

以戊二醛作为接枝改性剂制备改性骨胶粘合剂。研究了醛胶比、接枝温度和接枝时间等对改性骨胶粘合剂性能的影响,并采用红外光谱(FT-IR)和热失重分析(TGA)法对产物的结构与热稳定性进行了表征。结果表明:当醛胶比为0.2∶100、接枝温度为30℃和接枝时间为60 min时,所得改性骨胶粘合剂的剪切强度为10.7 MPa、凝胶温度为-2℃;此时产物的接枝效果明显,并具有良好的热稳定性能。     

人细小病毒B19的特性及其与血浆蛋白制品的相关性

人细小病毒B19(B19病毒)是一种直径约为20～25nm的无包膜单链线状DNA病毒,属于细小病毒家族。B19病毒对人类骨髓细胞,特别是红系祖细胞有高度趋向性,易感人群感染B19病毒可引起多种严重疾病,一般通过呼吸道和母婴垂直传播,但通过输注血液和血浆蛋白制品传播也非常普遍,已引起人们的广泛关注。本文对B19病毒的特性及其与血浆蛋白制品的相关性作一综述。